Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1998 Aug;78(3):328–335. doi: 10.1038/bjc.1998.494

Comparison of the cytotoxic activity of melphalan with L-prolyl-m-L-sarcolysyl-L-p-fluorophenylalanine in human tumour cell lines and primary cultures of tumour cells from patients.

R Larsson 1, S Dhar 1, H Ehrsson 1, P Nygren 1, R Lewensohn 1
PMCID: PMC2063025  PMID: 9703278

Abstract

m-L-sarcolysin (m-L-SL) is an isomer of melphalan (Mel) with the di(2-chloroethyl) amino group being substituted in the meta position of phenylalanine. By covalent conjugation of amino acids to m-L-SL, a peptide complex consisting of six m-L-SL-based oligopeptides known as peptichemio (PTC) was developed previously. In the present study, the cytotoxic activity pattern of the different oligopeptides of PTC was investigated in ten human tumour cell lines representing different mechanisms of cytotoxic drug resistance using the fluorometric microculture cytotoxicity assay (FMCA). In the cell line panel, L-prolyl-m-L-sarcolysyl-L-p-fluorophenylalanine (P2) was the most active oligopeptide, showing slightly lower mean IC50 values (2.6 vs 3.9 and 4.1 microg ml(-1)) than Mel and m-L-SL. The other five oligopeptides were less active than Mel. All active oligopeptides showed mechanistic similarity to Mel as judged by the correlation analysis of the cell line panel log IC50 values (R > or = 0.90), although P2 appeared to be less sensitive to GSH-mediated drug resistance. The relative activity of Mel and P2 was found to be related to degree of proliferation, P2 being more active towards low-proliferating cell lines. P2 and Mel were then further characterized in 49 fresh human tumour samples. In these samples P2 was considerably more active than Mel and showed a higher relative solid tumour activity (2.7 to 4.5-fold). However, the correlation of log IC50s between P2 and Mel in patient cells was high (R = 0.79), indicating a similar mechanism of action in this tumour model too. Cross-resistance with other standard drugs was lower for P2 than Mel. The results show that P2 is the most potent component of PTC and demonstrates a favourable activity profile compared with Mel. These data suggest that further investigation of P2 as a potential anti-tumour agent is warranted.

Full text

PDF
328

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Batist G., Torres-Garcia S., Demuys J. M., Greene D., Lehnert S., Rochon M., Panasci L. Enhanced DNA cross-link removal: the apparent mechanism of resistance in a clinically relevant melphalan-resistant human breast cancer cell line. Mol Pharmacol. 1989 Aug;36(2):224–230. [PubMed] [Google Scholar]
  2. Bellamy W. T., Dalton W. S., Gleason M. C., Grogan T. M., Trent J. M. Development and characterization of a melphalan-resistant human multiple myeloma cell line. Cancer Res. 1991 Feb 1;51(3):995–1002. [PubMed] [Google Scholar]
  3. Botling J., Liminga G., Larsson R., Nygren P., Nilsson K. Development of vincristine resistance and increased sensitivity to cyclosporin A and verapamil in the human U-937 lymphoma cell line without overexpression of the 170-kDa P-glycoprotein. Int J Cancer. 1994 Jul 15;58(2):269–274. doi: 10.1002/ijc.2910580221. [DOI] [PubMed] [Google Scholar]
  4. Cole S. P., Bhardwaj G., Gerlach J. H., Mackie J. E., Grant C. E., Almquist K. C., Stewart A. J., Kurz E. U., Duncan A. M., Deeley R. G. Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. Science. 1992 Dec 4;258(5088):1650–1654. doi: 10.1126/science.1360704. [DOI] [PubMed] [Google Scholar]
  5. Csoka K., Larsson R., Tholander B., Gerdin E., de la Torre M., Nygren P. Cytotoxic drug sensitivity testing of tumor cells from patients with ovarian carcinoma using the fluorometric microculture cytotoxicity assay (FMCA). Gynecol Oncol. 1994 Aug;54(2):163–170. doi: 10.1006/gyno.1994.1187. [DOI] [PubMed] [Google Scholar]
  6. Csoka K., Liliemark J., Larsson R., Nygren P. Evaluation of the cytotoxic activity of gemcitabine in primary cultures of tumor cells from patients with hematologic or solid tumors. Semin Oncol. 1995 Aug;22(4 Suppl 11):47–53. [PubMed] [Google Scholar]
  7. Dalton W. S., Durie B. G., Alberts D. S., Gerlach J. H., Cress A. E. Characterization of a new drug-resistant human myeloma cell line that expresses P-glycoprotein. Cancer Res. 1986 Oct;46(10):5125–5130. [PubMed] [Google Scholar]
  8. Danks M. K., Schmidt C. A., Cirtain M. C., Suttle D. P., Beck W. T. Altered catalytic activity of and DNA cleavage by DNA topoisomerase II from human leukemic cells selected for resistance to VM-26. Biochemistry. 1988 Nov 29;27(24):8861–8869. doi: 10.1021/bi00424a026. [DOI] [PubMed] [Google Scholar]
  9. Danks M. K., Yalowich J. C., Beck W. T. Atypical multiple drug resistance in a human leukemic cell line selected for resistance to teniposide (VM-26). Cancer Res. 1987 Mar 1;47(5):1297–1301. [PubMed] [Google Scholar]
  10. Dhar S., Nygren P., Csoka K., Botling J., Nilsson K., Larsson R. Anti-cancer drug characterisation using a human cell line panel representing defined types of drug resistance. Br J Cancer. 1996 Sep;74(6):888–896. doi: 10.1038/bjc.1996.453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ehrsson H., Lewensohn R., Wallin I., Hellström M., Merlini G., Johansson B. Pharmacokinetics of peptichemio in myeloma patients: release of m-L-sarcolysin in vivo and in vitro. Cancer Chemother Pharmacol. 1993;31(4):265–268. doi: 10.1007/BF00685669. [DOI] [PubMed] [Google Scholar]
  12. Fridborg H., Nygren P., Dhar S., Csoka K., Kristensen J., Larsson R. In vitro evaluation of new anticancer drugs, exemplified by vinorelbine, using the fluorometric microculture cytotoxicity assay on human tumor cell lines and patient biopsy cells. J Exp Ther Oncol. 1996 Sep;1(5):286–295. [PubMed] [Google Scholar]
  13. Gingold N., Pittermann E., Stacher A. Peptichemio in the therapy of malignancies (phase-I-study). The evaluation committee. Int J Clin Pharmacol. 1974 Oct;10(3):190–202. [PubMed] [Google Scholar]
  14. Hansson J., Lewensohn R., Ringborg U. Cytotoxicity and DNA cross-linking induced by peptide conjugated m-L-sarcolysin in human melanoma cells. Anticancer Res. 1991 Sep-Oct;11(5):1725–1730. [PubMed] [Google Scholar]
  15. Hug V., Hortobagyi G. N., Buzdar A. U., Blumenschein G. R., Grose W., Burgess M. A., Bodey G. P. A phase II study of peptichemio in advanced breast cancer. Cancer. 1980 May 15;45(10):2524–2528. doi: 10.1002/1097-0142(19800515)45:10<2524::aid-cncr2820451010>3.0.co;2-m. [DOI] [PubMed] [Google Scholar]
  16. Jonsson E., Fridborg H., Csóka K., Dhar S., Sundström C., Nygren P., Larsson R. Cytotoxic activity of topotecan in human tumour cell lines and primary cultures of human tumour cells from patients. Br J Cancer. 1997;76(2):211–219. doi: 10.1038/bjc.1997.364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Larsson R., Fridborg H., Liliemark J., Csoka K., Kristensen J., de la Torre M., Nygren P. In vitro activity of 2-chlorodeoxyadenosine (CdA) in primary cultures of human haematological and solid tumours. Eur J Cancer. 1994;30A(7):1022–1026. doi: 10.1016/0959-8049(94)90136-8. [DOI] [PubMed] [Google Scholar]
  18. Larsson R., Kristensen J., Sandberg C., Nygren P. Laboratory determination of chemotherapeutic drug resistance in tumor cells from patients with leukemia, using a fluorometric microculture cytotoxicity assay (FMCA). Int J Cancer. 1992 Jan 21;50(2):177–185. doi: 10.1002/ijc.2910500204. [DOI] [PubMed] [Google Scholar]
  19. Larsson R., Nygren P. Cytotoxic activity of topoisomerase II inhibitors in primary cultures of tumor cells from patients with human hematologic and solid tumors. Cancer. 1994 Nov 15;74(10):2857–2862. doi: 10.1002/1097-0142(19941115)74:10<2857::aid-cncr2820741019>3.0.co;2-5. [DOI] [PubMed] [Google Scholar]
  20. Lewensohn R., Ehrsson H., Hansson J., Ringborg U. Increased toxicity and DNA cross-linking by peptide bound m-L-sarcolysin (Peptichemio) as compared to melphalan and m-L-sarcolysin in human melanoma cell lines. Anticancer Res. 1991 Jan-Feb;11(1):321–324. [PubMed] [Google Scholar]
  21. Lewensohn R., Fernberg J. O., Ehrsson H., Merlini G. Efficacy of peptide bound m-L-sarcolysin (peptichemio) on melphalan resistant human myeloma cells in vitro. Med Oncol Tumor Pharmacother. 1991;8(4):265–269. doi: 10.1007/BF02987196. [DOI] [PubMed] [Google Scholar]
  22. Merlini G., Gobbi P. G., Riccardi A., Riva G., Sardi C., Perugini S. Peptichemio induction therapy in myelomatosis. Cancer Chemother Pharmacol. 1982;8(1):9–16. doi: 10.1007/BF00292864. [DOI] [PubMed] [Google Scholar]
  23. Mirski S. E., Gerlach J. H., Cole S. P. Multidrug resistance in a human small cell lung cancer cell line selected in adriamycin. Cancer Res. 1987 May 15;47(10):2594–2598. [PubMed] [Google Scholar]
  24. Mulcahy R. T., Bailey H. H., Gipp J. J. Up-regulation of gamma-glutamylcysteine synthetase activity in melphalan-resistant human multiple myeloma cells expressing increased glutathione levels. Cancer Chemother Pharmacol. 1994;34(1):67–71. doi: 10.1007/BF00686114. [DOI] [PubMed] [Google Scholar]
  25. Nygren P., Csoka K., Jonsson B., Fridborg H., Bergh J., Hagberg H., Glimelius B., Brodin O., Tholander B., Kreuger A. The cytotoxic activity of Taxol in primary cultures of tumour cells from patients is partly mediated by Cremophor EL. Br J Cancer. 1995 Mar;71(3):478–481. doi: 10.1038/bjc.1995.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nygren P., Fridborg H., Csoka K., Sundström C., de la Torre M., Kristensen J., Bergh J., Hagberg H., Glimelius B., Rastad J. Detection of tumor-specific cytotoxic drug activity in vitro using the fluorometric microculture cytotoxicity assay and primary cultures of tumor cells from patients. Int J Cancer. 1994 Mar 1;56(5):715–720. doi: 10.1002/ijc.2910560517. [DOI] [PubMed] [Google Scholar]
  27. Nygren P., Kristensen J., Jonsson B., Sundström C., Lönnerholm G., Kreuger A., Larsson R. Feasibility of the fluorometric microculture cytotoxicity assay (FMCA) for cytotoxic drug sensitivity testing of tumor cells from patients with acute lymphoblastic leukemia. Leukemia. 1992 Nov;6(11):1121–1128. [PubMed] [Google Scholar]
  28. Nygren P., Larsson R. Verapamil and cyclosporin A sensitize human kidney tumor cells to vincristine in absence of membrane P-glycoprotein and without apparent changes in the cytoplasmic free Ca2+ concentration. Biosci Rep. 1990 Apr;10(2):231–237. doi: 10.1007/BF01116583. [DOI] [PubMed] [Google Scholar]
  29. Paccagnella A., Salvagno L., Chiarion-Sileni V., Bolzonella S., De Besi P., Frizzarin M., Pappagallo G. L., Fosser V. P., Fornasiero A., Segati R. Peptichemio in pretreated patients with plasmacell neoplasms. Eur J Cancer Clin Oncol. 1986 Sep;22(9):1053–1058. doi: 10.1016/0277-5379(86)90005-2. [DOI] [PubMed] [Google Scholar]
  30. Paccagnella A., Tredese F., Salvagno L., Brandes A., Sileni V. C., Daniele O., Fornasiero A., Fosser V., Nicoletto O., Maggino T. Peptichemio in pretreated patients with ovarian cancer. Cancer Treat Rep. 1985 Jan;69(1):17–20. [PubMed] [Google Scholar]
  31. STERN K., BIRMINGHAM M. K., CULLEN A., RICHER R. Peptidase activity in leucocytes, erythrocytes and plasma of young adult and senile subjects. J Clin Invest. 1951 Jan;30(1):84–89. doi: 10.1172/JCI102420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yagi M. J., Bekesi J. G., Daniel M. D., Holland J. F., De Barbieri A. Increased cancericidal activity of PTT.119, a new synthetic bis-(2-chloroethyl)amino-L-phenylalanine derivative with carrier amino acids. I. In vitro cytotoxicity. Cancer Chemother Pharmacol. 1984;12(2):70–76. doi: 10.1007/BF00254592. [DOI] [PubMed] [Google Scholar]
  33. Yagi M. J., Scanlon K. J., Chin S. E., Holland J. F., Bekesi J. G. Multiple transport pathways for L1210 cells: uptake of PTT.119, a bifunctional alkylator with carrier amino acids. Chemotherapy. 1988;34(3):235–247. doi: 10.1159/000238575. [DOI] [PubMed] [Google Scholar]
  34. Zaniboni A., Simoncini E., Marpicati P., Montini E., Rossi G., Marini G. Peptichemio, teniposide and high-dose dexamethasone: a new active combination for relapsing and refractory multiple myeloma. A pilot study. Anticancer Res. 1988 Jan-Feb;8(1):125–127. [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES